Herbal compositions and treatment methods

ABSTRACT

An herbal-based treatment for Hepatitis C includes boiling a filtered residue from the  Ecballium Elaterium  plant to form a condensate. This condensate is mixed with water and administered in the form of drops. The drops were given to human patients, along with a single herb, Camelthorn, in a powder form. The combined herbal-based treatment was administered to human beings who were chronically infected with Hepatitis C. These patients, in terms of reduction of viral loads, normalization of enzymes, and general well being, were treated successfully in most instances.

BACKGROUND OF INVENTION

[0001] Hepatitis is a disease of the liver, which is usually caused byviral infections. A significant number of patients suffering fromhepatitis are infected with either hepatitis B virus (HBV) or hepatitisC virus (HCV). HCV is an RNA virus with a virion size of 30-60 nm. HBVis a DNA virus with a virion size of 42 nm (D. S. Chen, J. Formos. Med.Assoc., 95(1):6-12 (1996)). Serious complications of hepatitis caninclude cirrhosis, hepatic failure, liver cancer and death.

[0002] As of 1997, nearly 14 million people in the United States and 100million worldwide were infected with the hepatitis C virus (Alter, M.J., Hepatology, 26(1):625-655 (1997)). It is expected that worldwide,the number of patients has increased to 170 million people in 2002(ViroPharma Inc. Press Release, Jun. 19, 2002).

[0003] Chronic hepatitis C is the most common cause of chronic liverdisease and is the leading indication for liver transplantation in theUnited States (Detre, et al., Viral Hepatitis Rev., 2:219-228 (1996)).

[0004] In Egypt, HCV represents the highest donor infection ratesrecorded so far where 14-24% of blood donors are anti-HCV positive(Hipps, et al., Infect. Dis. 168:789-790 (1993)).

[0005] Hepatitis B is a major health problem worldwide, particularly inAsia and Africa. Approximately 300 million people are chronicallyinfected with HBV worldwide and more than one million carriers of HBVare found in the United States. HBV carriers not only become long-termreservoirs of the virus, but also may develop chronic liver disease andhave a greatly increased risk of developing liver cirrhosis and cancer.

[0006] Unfortunately, there are few effective treatments for hepatitis.Treatment of HCV with recombinant-alpha interferon has a limitedlong-term efficacy with a response rate of only about 25% (D. S. Chen,J. Formos. Med. Assoc., 95(1):6-12 (1996)). A long-term relapse rate of24% was reported in Chinese patients with chronic hepatitis B whounderwent interferon therapy (Lok, et al., Gastroenterology,105(6):1833-1838 (1993)). Furthermore, interferon is associated with anumber of adverse side effects, including thrombocytopenia, leucopenia,bacterial infections, and influenza-like symptoms. Other agents used inthe treatment of hepatitis include the nucleoside analog ribavirin andursodeoxycholic acid. However, neither has been shown to be particularlyeffective (D. C. Dale and D. D. Federman, eds. Medicine Sc. Amer., Inc.,New York, 4(8):1-8 (1995)). The most effective treatment now in widespread use may be the combination of Interferon and Ribavirin.(McHutchison et at New England Journal of Medicine, Vol 339, pps.1485-1491.

[0007] The use of herbal therapy and folk medicines has been known forthousands of years in China. In fact, records on the use of herbs dateback to biblical times. However, only recently have scientists begunexploring the possible role for herbs in treatment of viral infections.For example, extracts of Hemidesmus indicus rhizomes (Family:Asclepiadaceae) have been found to have anti-viral activity against DNAand RNA viruses (Babbar, et al., Ind. J. Exp. Biol., 8:304-312 (1970)).Likewise, extracts from the root of the Ecballium Elaterium have beenused to treat HCV and HBV (EP 0793964 and U.S. Pat. No. 5,648,089).While research in the field of herbal medicines has increased, muchremains to be learned about the effectiveness of such herbal remedies.

[0008] In view of the above, there is a continuing need in the art foralternative options for treating viral infections such as HCV and HBV.Moreover, there is a need in the art for treatment options that provideminimal side effects.

[0009] Accordingly, it is an object of the present invention to provideherbal compositions and methods for treating viral infections such asHCV and HBV. Moreover, it is object of the present invention to providecompositions and methods for treatment that incur minimal side effectsto the subject. Another object of the invention is to develop atreatment, which involves a minimum of only two herbal elements. Thislast object is of vital importance to the eventual large-scalecommercial use of a herbal treatment. Current United States regulationsimpose strict manufacturing controls on the manufacture ifpharmaceutical products. Herbal treatments based upon too many herbalcomponent compound manufacturing processes and may make these morecomplicated treatments commercially impracticable.

[0010] The inventors have previously developed a treatment for bothHepatitis C and Hepatitis B, along with other virus caused diseases. Theprior attempts involved many different herbs. Some were thought to beanti-viral and some had the purpose of reducing or eliminating sideeffects. Each treatment had at least ten herbs or extracts from herbs.After a study of United States Federal Drug Administration proposedregulations, the inventors sought to come up with a treatment in whichonly one or two herbs or extracts from herbs would be used.

[0011] One of the extracts was from the herb, Ecballium Elaterium. Thisherb had long been used to treat liver disease such as jaundice. (See,“Medicinal Plants of North Africa”, Loutfy Boulos, ReferencePublications, Inc 1983, page ______) Ecballium Elaterium could not beused directly as it has some serious toxicity problems.

[0012] See A Manual of Materia Medica and Pharmacology by David M. R.Culbreth, Ph. G., M.D. (1927) which describes the properties of EcbaliumElaterium as follows: “PROPERTIES—Hydragogue cathartic (most powerfulknown), producing profuse watery evacuations with griping and muchprostration; large doses nauseate, vomit, inflame stomach and bowels,increase flow of urine, and may kill. Does neither vomit nor purge dogs,rabbits, but kills them by convulsions. Those working in it often haveulcerated fingers, eyes, etc”.

[0013] Initially the inventors obtained an extract of Ecbalium elateriumpreparing herbal drops made by using the fruit of Ecbalium elaterium andwashing the fruit with clean water. Approximately a half-kilogram offruit and one liter of distilled water were combined in an ordinaryhousehold electrical grinder. The fruit was ground for two minutes. Themixture was then poured through a sieve of cotton gauze to separate outthe larger fruit particles. The mixture was then filtered using Wattmannfilter paper. The filtrate (i.e., cell sap) was stored in a refrigeratorat 10° C. for approximately 5-7 hours to promote the precipitation ofmicroparticles. The filtrate was then filtered using ceramicsterilization candles to remove any microorganisms. The pH was adjustedto approximately 6.8-7 using dilute sulfuric acid and sodiumbicarbonate. A preservative (thiomersal) was added to the sterilizedfiltrate to provide a concentration of 0.001 gm/liter. The herbalsolution was then placed into 10 cc. dropper bottles under asepticconditions and stored in the refrigerator at about 4 degrees C. Thefinal concentration of the extract was approximately 0.5 weight % cellsap based on the amount of cell sap dissolved in water (5 gms/1000 ml).

[0014] This method did produce a successful anti-viral but those treatedwith Ecbalium elaterium did require additional herbs to minimize sideeffects. As a result, the treatment had too many herbs, making itsfuture manufacture unfeasible.

[0015] The inventors noticed that after placing aluminum foil over theEcbalium elaterium mixture (in order to protect it from dust) a brownresidue would form on the foil. They then isolated the residue andreduced it to liquid form so as to be administered in drops. Theinventors surmised that the Ecbalium evaporate might contain ananti-viral and additionally might not contain the toxic part of Ecbaliumelaterium. They thought that the evaporate was present at 40 degrees C.,as this was roughly the summer temperature in which the Ecbaliumelaterium was stored and the discoloring of the covering aluminumoccurred. After discarding the use of water, methyl alcohol, and ethylalcohol, the inventors tried methylene chloride. Methylene chloride isnot soluble in water, has mild polarity, and boiling point of 30 to 32degrees C.

[0016] The inventors add the same volume of the solvent, methylenechloride, and the yield-differentiated distillation (using tap water andfresh Ecbalium fruit), then hand shaking for about five minutes in aseparating funnel, then leaving the mixture for 30 minutes. This laststep gives the solvent a chance for the solvent to be separated form thewater due to the different densities. The solvent, which contained theEcbalium elaterium residue, is then transferred to a rotary evaporationmachine. The machine evaporates the solvent away at 29 to 30 degrees C.,under vacuum. This process yields about a half milliliter of residueover a period of 3 to 4 hours. The residue is a strong smelling brownoily substance.

[0017] A HPLC machine analyzed the above final residue. The result wasthe largest indication of absorption at 8.648 AU (Angstrom) but thesubstance remains chemically unidentified. The mechanism of action maybe similar to that found by others in extract from Aloe Vera. (See U.S.Pat. No. 5,118,673 Carpenter, et al. Jun. 2, 1992 Uses of aloe products)

[0018] The inventors, using their wide experience in herbal therapy,surmised that the herb Camel thorn (Alhagi pseudalhagi), with its longrecord of versatility, would be a good candidate to test as ananti-viral. Their trial dose of Camel thorn (Alhagi pseudalhagi) inpowder form was one teaspoon-full twice a day before meals. Theysubsequently prepared the single herb in the form of drops by steamdistillation process by boiling 0.5 kg of cleaned shoots in 2 liter oftap water thereby gaining 500 ml of a mixture of distilled water andvapor materials. Then, the drops are packed into 10-ml plastic vials.The dose of single herb Camel thorn (Alhagi pseudalhagi) drops is 4drops nasally once a day.

[0019] Since it was known that a combination of ingredients can be moreeffective in treating Hepatitis C (see Interferon Alfa-2b Alone or incombination with Ribavirin as Initial Treatment for Chronic Hepatitis C,McHutchison et al, New England Journal of Medicine, Vol 339, p1485-1491, Nov. 19, 1998), the inventors tested both the herbs Alhagipseudalhagi and Ecballium elaterium residue both singly and incombination.

SUMMARY OF THE INVENTION

[0020] The present invention provides three herbal compositions havinganti-viral properties and methods of treating viral infections. Theseare:

[0021] 1. An extract of Ecballium elaterium (squirting cucumber)prepared in a unique manner.

[0022] 2. A single herb, Alhagi pseudalhagi (camel thorn).

[0023] 3. An extract of Ecballium elaterium, prepared in a uniquemanner, and a single herb, Alhagi pseudalhagi.

[0024] The present invention also provides a method for treating a viralinfection, which includes administering to a subject an effective amountof either of the three above herbal compositions being selected fromamong the herbal compositions of the invention. The subjects to betreated are preferably subjects in need of treatment (suffering from aviral infection). Subjects to be treated are preferably mammals inparticular humans. Viral infections to be treated include hepatitis C,hepatitis B or a combination of the two and other viral infections suchas influenza, viral diabetes, and common cold.

[0025] Advantageously, the herbal compositions and methods of thepresent invention provide an alternative treatment option for subjectssuffering from viral infections. In addition, the herbal compositionsand methods of the present invention provide an alternative treatmentoption that exhibits minimal side effects. These and other advantages ofthe present invention will become readily apparent from the detaileddescription set forth below.

[0026] In the context of the invention, “treating” or “treatment” at aminimum refers to inhibiting the progression of the viral infection,which can be ascertained qualitatively (e.g., by a reduction in clinicalsymptoms) or quantitatively (e.g., by a reduction in viral load or otherquantifiable criteria.). An “effective” amount is any amount of theherbal composition that inhibits or stops the progression of the viralinfection. However, as will be apparent to those skilled in the art, theefficacy of a particular composition on a subject will be affected by avariety of factors including, but not limited to, the method ofadministration, the body mass and age of the subject, and the stage ofthe infection (e.g., acute or chronic).

[0027] As previously described, the herbal compositions of the presentinvention are particularly suitable for treating a subject having aviral infection. Viruses to be treated include DNA and RNA virus, withhepatitis B virus (HBV) and the hepatitis C virus (HCV) beingparticularly preferred. In the treatment of HBV or HCV, inhibition ofinfection can be qualitatively ascertained by determining the presenceor absence of the viral genome by polymerase chain reaction (PCR) andreverse transcriptase polymerase chain reaction (RT-PCR), or byobserving a reduction and/or disappearance in clinical symptoms such asjaundice, weakness, right hypochondrial pain, loss of appetite, andother symptoms that are associated with hepatitis infection. Efficacy intreating of HBV or HCV can also be quantitatively ascertained bymeasuring a reduction in viral load using PCR and RT-PCR or by measuringa reduction (i.e., normalization) in elevated liver enzyme levels (e.g.,serum alanine transaminase (ALT) and serum aspartate transaminase(AST)). Alternatively, assays for surface antigens of HBV (e.g., HBsAg)and HCV can also be used.

[0028] Subjects to whom the herbal compositions of the invention areadministered include any living organism, with mammals such as primatesand humans being more preferred. The subject is also preferably in needof treatment (i.e., has a viral infection that requires treatment).Accordingly, in a more preferred embodiment, the subject is infectedwith HBV, HCV or both.

[0029] The herbal compositions are administered to the subject by anytechnique known in the art. Routes of delivery can include, but are notlimited to, oral, intranasal, sublingual, intrapulmonary, rectal,transdermal, parenteral and combinations thereof. Acceptable dosageforms suitable for administration to a subject include, but are notlimited to, tablets, capsules, powders, patches, solutions, andsuspensions. The compositions of the invention can include aphysiologically-acceptable carrier in which the herbal extracts aredispersed. For example, the carrier can be buffered saline if a liquiddosage unit is to be prepared. Procedures for making and administeringsuch dosage forms are well within the abilities of one of ordinary skillin the art.

[0030] In one particular embodiment, the subject is administered aneffective amount of the compositions of the invention, The combined useof these two herbal compositions have been found to be particularlyeffective in treating subjects infected with HBV and HCV, as compared tousing the compositions alone.

DETAILED DESCRIPTION OF THE INVENTION

[0031] In accordance with the present invention there are providedherbal compositions useful for the treatment of viral infections, suchas hepatitis, and methods of using these compositions. The herbalcompositions of the invention are particularly useful for the treatmentof hepatitis B and hepatitis C.

[0032] Subjects to whom the herbal compositions of the invention areadministered include any living organism, with mammals such as primatesand humans being more preferred. The subject is also preferably in needof treatment (i.e., has a viral infection that requires treatment).Accordingly, in a more preferred embodiment, the subject is infectedwith HBV, HCV or both.

[0033] The herbal compositions are administered to the subject by anytechnique known in the art. Routes of delivery can include, but are notlimited to, oral, intranasal, sublingual, intrapulmonary, rectal,transdermal, parenteral and combinations thereof. Acceptable dosageforms suitable for administration to a subject include, but are notlimited to, tablets, capsules, powders, patches, solutions, andsuspensions. The compositions of the invention can include aphysiologically-acceptable carrier in which the herbal extracts aredispersed. For example, the carrier can be buffered saline if a liquiddosage unit is to be prepared. Procedures for making and administeringsuch dosage forms are well within the abilities of one of ordinary skillin the art.

[0034] In one particular embodiment, the subject is administered aneffective amount of the compositions of the invention, with thetwo-component herbal composition being preferred as the primary herbalcomposition. The primary herbal composition is preferably administeredin liquid form and in a dosage of 2 drops, two times per day. Asecondary herbal composition, the Ecbalium elaterium residue only isadditionally administered to the subject orally in a dosage of twodrops, two times a day. In another embodiment, the liquid drops orpowder consisting only of Camel thorn (Alhagi pseudalhagi) areadministered either liquid, 2 drops twice a day, or powder, one to fivegrams, three times a day, with 1 to 3 grams, three times a day beingpreferred. The combined use of these two herbal compositions have beenfound to be particularly effective in treating subjects infected withHBV and HCV, as compared to using the compositions alone.

[0035] Extracts of the various herbs are prepared following conventionaltechniques known in the art. The extracts can be prepared as a powderedextract using a maceration extraction process or as a liquid extractusing a solvent extraction process. For example, the herbal compositionsmay be prepared as herbal extracts as taught in U.S. Pat. No. 5,876,728,or in U.S. Pat. No. 5,874,804

[0036] The herbal compositions are not limited to any one particulardosage form. The compositions of this invention can be prepared in avariety of dosage forms known in the art. However, as will be apparentto those skilled in the art, the particular dosage form used will bedependent on the delivery route. The herbal compositions can alsoinclude excipients to alter taste, bulk, and texture, and can includepreservatives to increase shelf life.

[0037] In accordance with the present invention, methods of treatingviral infections with the herbal compositions of the invention are alsoprovided. The methods entail administering to the subject an effectiveamount of a primary herbal composition, which includes:

HCV Study

[0038] To determine the efficacy of the herbal compositions of theinvention, an open 18-month study was conducted with 23 patientsinfected with the hepatitis C virus. The study involved a treatmentregimen using the herbal compositions of the three herbal compositionsdescribed above.

[0039] Criteria for entry into the study were as follows: (a) testingserum-positive for HCV antibodies within the last 6 months before entryto the study; (b) testing serum-positive for HCV-RNA by RT-PCR withinthe last 3 months before entry to the study; (c) exhibiting elevatedlevels of serum alanine transaminase (ALT) and serum aspartatetransaminase (AST) within the last 6 months before entry into the study;(d) exhibiting ALT levels at least twice the upper normal level withinthe last 6 months before entry into the study which at no time duringthis time period dropped to or below the normal level, and (e) testedagain for HCV-RNA by RT-PCR. Patients were excluded from the study ifthey had another major illness such as major active infections, cancer,or renal failure; evidence of liver disease other than viral hepatitis;or a known history or presence of ascites, hepatomegaly or livercirrhosis. Patients were also excluded if they were alcoholic or a drugabuser, concurrently used other herbs or folk medicine, concurrentlyused other anti-viral medication, or demonstrated other unsuitablecharacteristics including severe allergies or dizziness. Dizziness wasconsidered as a criterion for excluding patients because it occurredoccasionally after taking the herbal compositions.

[0040] RT-PCR of serum was done following conventional techniques usinga Sorine kit. RT-PCR was followed by thirty-five (35) cycles ofamplification of HCV cDNA. A Triple test (El-Awady, et al., Chem.Clinical Acta, 283(1-2):1-14 (1999)) was also conducted for somepatients to assay HCV-RNA in serum and leucocytes (both positive andnegative strands). In accordance with this test, negative and positivecontrols were run with each assay to avoid false negatives and falsepositives. In addition, as part of the Triple test, subjects were testedfor serum viraemia, mature viral genome in peripheral mononuclear celllysate as well as replicating forms of virus in peripheral leucocytes.This gave an impression of the extra hepatic tissue, in which theabsence of replicating forms in extra hepatic tissue after treatmentprovided a good prognosis of the effectiveness of the herbalcompositions. Other PCR tests such b-DNA and AMP were used as well.

[0041] The serum AST and ALT levels were assayed following knowntechniques (Bergmeyer and Horder, Clin. Chen. Acta 105147 f. (1980);International Federation of Clinical Chemistry, Scientific Committee, J.Clin. Chem. Biochem., 18:521-534 (1980); Bergmeyer H. U. Principles ofEnzymatic Analysis. Verlag Chemic, (1978)). Assay kits manufactured byStanbio Laboratories, Inc., (USA), Bio. Merieux (France), and BoehringerMannheim (Germany) were used.

[0042] The patients were divided into 3 groups. The first group (GroupA) received the single herb, Alhagi pseudalhagi. The second group (GroupB) received herbal drops of the Ecaballium elaterium residue. The thirdgroup (Group C) received the single herb Alhagi pseudalhagi and theherbal drops made from the Ecaballium Elaterium residue. (Table 1.)

[0043] The health status of each patient was monitored monthly, whichincluded monitoring changes in body weight, blood pressure, eatinghabits, sleeping habits, strength, pain, and overall appearance. SerumAST and ALT levels were also monitored monthly in addition to otherenzyme levels. Following generally accepted practices, normalization ofAST and ALT levels was considered to occur when levels fell below twicethe normal limits.

[0044] Generally, RT-PCR of patients' serum was additionally performedafter 6 months of treatment. (Table 2.)

[0045] The percentage of patients having normal enzyme levels (i.e.,less than twice the normal limit) at the beginning of treatment andduring the first 4 months of treatment is listed in Table 3. After 4months, the number of patients that returned for monthly assessment ofenzyme levels began to significantly decrease, TABLE 1 Grouping ofPatients Treatment Group A Single herb Group B Herbal drops Group CSingle herb + herbal drops

[0046] TABLE 2 PCR readings for each patient after treatment GroupPatients High Moderate Weak Negative A 4 1 3 0 0 B 12 2 4 3 3 C 23 1 4 87

[0047] TABLE 3 Enzyme readings for each patient after treatment SPGTLevels Group Patients Normalized Elevated A 4 3 1 B 12 12 0 C 23 18 5

[0048] Furthermore, all patients in all Groups reported an improvementin symptoms associated with hepatitis such as:

[0049] 1. Increase in strength sufficient to allow patient workefficiently;

[0050] 2. Disappearance of right hypochondrial pain and so the patientcan sleep on his right side;

[0051] 3. An improvement of prothrombine synthesis and a correspondingdecrease in bleeding;

[0052] 4. Healing of skin lesions, disappearance of tenderness of skinand soft tissues, and clearance of skin pigmentations in hepaticpatients;

[0053] 5. Improvement of appetite and digestion;

[0054] 6. Loss of constipation and correction of bowel habits;

[0055] 7. Disappearance of an earthy look of the face;

[0056] 8. Decrease attacks of cholecystitis; and

[0057] 9. Weight gain.

We claim:
 1. A herbal composition, which comprises: (a) A single herb;and (b) A residue derived from an evaporate substance of fruit ofEcballium Elaterium. (c) A combination of a single herb and a residuederived from an evaporate substance of the fruit of Ecballium Elaterium.2. A method for treating a viral infection, which comprisesadministering to a subject an effective amount of a primary herbalcomposition, which comprises: (a) a single herb; and (d) a residuederived from an evaporate of fruit of Ecballium Elaterium. (e) Acombination of a single herb and a residue derived from an evaporate ofthe fruit of Ecballium Elaterium.
 3. The method according to claim 2,wherein the viral infection is hepatitis B, Hepatitis C or a combinationthere of.
 4. The method according to claim 2, wherein the subject is amammal
 5. The method according to claim 2, wherein the mammal is ahuman.
 6. The method according to claim 2, wherein the subject is in theneed of such treatment.